FOXA1/HNF3A

FOXA1/HNF3A CUTANA™ CUT&RUN Antibody

$445.00
SKU: 13-2001
Pack size: 100 µL
  • Type:  Polyclonal
  • Host:  Rabbit
  • Target Size:  49 kDa
  • Format:  Affinity Purified IgG
  • Reactivity:  Human, Mouse (Predicted Rat)
  • Applications: CUT&RUN, WB, IP

Description

This antibody meets EpiCypher’s “CUTANA Compatible” criteria for performance in Cleavage Under Targets and Release Using Nuclease (CUT&RUN) and/or Cleavage Under Targets and Tagmentation (CUT&Tag) approaches to genomic mapping. Every lot of a CUTANA Compatible antibody is tested in the indicated CUTANA approach using EpiCypher optimized protocols and determined to yield peaks that show a genomic distribution pattern consistent with reported function(s) of the target protein. FOXA1 antibody produces CUT&RUN peaks above background primarily in intronic, intergenic, and promoter regions (Figure 1) that overlap with known FOXA1 DNA-binding motifs (Figure 2).

Validation Data

Figure 1: FOXA1 peaks in CUT&RUN
CUT&RUN was performed using 500,000 MCF7 cells with 0.5 µg FOXA1 antibody. Peaks were called using MACS2. (A) Heatmap showing FOXA1 peaks relative to IgG negative control antibody (EpiCypher 13-0042) in aligned rows ranked by intensity (top to bottom) and colored such that red indicates high localized enrichment and blue denotes background signal. (B) The number of peaks which fall into distinct classes of functionally annotated genomic regions is plotted.

Figure 2: FOXA1 transcription factor binding motif analysis in CUT&RUN
(A) Homer software was used to identify known motifs underneath FOXA1 CUT&RUN peaks. The FOXA1 consensus motif was highly enriched, and is represented as a sequence logo position weight matrix. (B) The number of FOXA1 peaks containing FOXA1 consensus motifs from panel A is represented by a Venn Diagram. (C) Three representative loci showing FOXA1 peaks overlapping with FOXA1 consensus motifs (Integrative Genomics Viewer).

Figure 3: Western blot detection of human and mouse FOXA1
Whole cell lysates were isolated from MCF-7 and HeLa cells using NETN lysis buffer. The indicated amounts (µg) of lysate were loaded onto 4-20% SDS-PAGE gel and analyzed under standard western blot conditions using FOXA1 antibody (0.1 µg/mL).

Figure 4: Immunoprecipitation of human FOXA1
EpiCypher FOXA1 antibody (6 µg) was used to immunoprecipitate whole cell lysates isolated from HeLa cells using NETN lysis buffer (1 mg per IP). A negative control IgG antibody and positive control antibody to a different FOXA1 epitope (Bethyl Laboratories) were also used to demonstrate specificity of the IP. Immunoprecipitates were loaded onto 4-20% SDS-PAGE gel (20% of IP loaded) and probed via western blot with EpiCypher FOXA1 antibody (1 µg/mL).

Technical Information

Immunogen
A synthetic peptide corresponding to human FOXA1 amino acids 422 to 472.
Formulation
Antigen affinity-purified antibody (1.0 mg/mL) in Tris-citrate/phosphate buffer pH 7 to 8, 0.09% sodium azide.
Storage and Stability
Stable for 1 year at 4°C from date of receipt.

Application Notes

Recommended Dilutions:

CUT&RUN: 0.5 µg

Western Blot (WB): 1:1,000 - 1:10,000

Immunoprecipitation (IP): 2 - 10 µg/mg lysate

Documents & Resources

Additional Info

This product is provided for commercial sale under license from Bethyl Laboratories, Inc.

Applications Key:

ChIP: Chromatin immunoprecipitation
CUT&RUN: Cleavage Under Targets and Release Using Nuclease
CUT&Tag: Cleavage Under Targets and Tagmentation
E: ELISA
FACS: Flow cytometry
ICC: Immunocytochemistry
IF: Immunofluorescence
IHC: Immunohistochemistry
IP: Immunoprecipitation
L: Luminex
WB: Western Blot

Reactivity Key:

B: Bovine
Ce: C. elegans
Ch: Chicken
Dm: Drosophila
Eu: Eukaryote
H: Human
M: Mouse
Ma: Mammal
R: Rat
Sc: S. cerevisiae
Sp: S. pombe
WR: Wide Range (predicted)
X: Xenopus
Z: Zebrafish
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